Role of TGFbeta in Microtubule Dynamics

TGFbeta 在微管动力学中的作用

基本信息

批准号：
7915833
负责人：
Kathleen M Mulder
金额：
$ 34.93万
依托单位：
PENNSYLVANIA STATE UNIV HERSHEY MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-01 至 2011-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7915833
关键词：
Affect Antineoplastic Agents Arts Back Binding Biological Cancer Patient Cell Cycle Inhibition Cell Cycle Progression Cell Nucleus Cells Characteristics Complex Cyclic AMP-Dependent Protein Kinases DNA Synthesis Inhibition DNA biosynthesis Data Defect Development Disease Dyes Dynein ATPase Early Endosome Endocytosis Endosomes Event Family Fibronectins Figs - dietary Future Gene Targeting Goat Growth Factor Human Image Immunoglobulin G Impairment In Vitro Incubated Intracellular Transport Investigation Left Ligand Binding Light Link MDCK cell Malignant Neoplasms Malignant neoplasm of ovary Mediating Microscope Microscopy Microtubules Modeling Motor Mus Mutate Mutation Nature Normal tissue morphology Nuclear Nuclear Translocation Organelles Oryctolagus cuniculus Pathway interactions Phosphorylation Phosphorylation Site Phosphotransferases Plasmids Play Preparation Protein Isoforms Proteins Recruitment Activity Regulation Reporting Resistance Role Serum Signal Transduction Signaling Molecule Small Interfering RNA Specific qualifier value Specificity Students System Testing Thymidine Time Tissues Transcriptional Regulation Transfection Transforming Growth Factor beta Transmembrane Transport Transport Vesicles Vesicle Western Blotting base cancer cell cell growth drug development dynein light chain functional outcomes in vivo innovation knock-down malignant phenotype member mutant neoplastic cell novel oncology receptor research study response trafficking tumor tumor progression

项目摘要

DESCRIPTION (provided by applicant): A higher order of regulation can be achieved by compartmentalization and functional specialization of the endocytic pathway during signal transduction in response to growth factors (GFs). In contrast, impairment of intracellular (IC) trafficking of GF signaling complexes can result in mis-localization or accumulation of signaling molecules in the wrong compartment, or at the wrong time, giving rise to various disease states, including cancer. In addition, the specificity of motor protein light chains (LCs) in recruiting their membranous cargo for IC transport during regulation of GF signaling is becoming increasingly clear. Additional investigation of the regulation, key components, timing, and functional outcome of signaling from endosomal subcompartments is required to enable oncology drug development, based upon these newer signaling regimes. Our data from the last project period indicated that km23 dynein LCs are activated by TGF? receptors (T?Rs) and are required for Smad trafficking, prior to nuclear translocation. Thus, km23 appears to function as a "motor receptor" to recruit TGF? signaling complexes to the dynein motor for IC transport of cargo (ie, T?Rs, Smads) along microtubules (MTs) toward the nucleus. In addition, we have identified another related member of the km23/LC7/robl/DYNLRB family, termed km23-2. While this isoform appears to have some characteristics similar to km23-1, km23-2 appears to more specifically regulate Smad3, in contrast to Smad2 regulation by km23-1. The mechanisms underlying the differential regulation and trafficking of Smads2/3 in relation to the km23-1/2 motor LCs will be explored in the new application, including how km23 impacts downstream TGF? responses. During the last project period, we have also shown that protein kinase A (PKA) directly phosphorylates km23 and is required for km23 binding to the dynein intermediate chain (DIC). The mechanisms underlying this PKA phosphorylation of km23 in TGF? signaling and Smad2/3 trafficking will also be examined. Additional studies relate to the investigation of how these trafficking events are altered in human ovarian cancer cells (HOCCs) known to express altered forms of km23-1. As part of the last project period, we identified km23-1 alterations in 42% of ovarian cancer patient tissues, with no km23 alterations detectable in normal tissues. Such a high alteration rate in ovarian cancer suggests that km23 may play an important role in either TGF2 resistance or tumor progression of this disease. In the new application, we will examine the effects of knocking down km23, or mutating it at key phosphorylation sites, on Smad trafficking, Smad-specific TGF? responses, and the malignant phenotype of the human cancer cells in vitro, as well as on tumor progression in vivo.

描述（由申请人提供）：通过响应生长因子（GFS）的信号转导期间，可以通过对内吞途径的隔室化和功能专业化来实现更高的调节。相比之下，GF信号传导复合物的细胞内（IC）运输受损可能会导致信号分子在错误的隔室中或错误的时间的错误定位或积累，从而导致包括癌症在内的各种疾病状态。另外，在调节GF信号调节期间，运动蛋白光链（LC）在募集其膜货物以进行IC转运方面的特异性变得越来越清晰。基于这些新的信号传导状态，需要对内体子组门的调节，关键组成部分，定时和功能结果进行进一步研究，以实现肿瘤药物的开发。我们从上一个项目期间开始的数据表明，KM23 Dynein LCS被TGF激活？受体（t？rs），在核易位之前需要SMAD贩运。因此，KM23似乎是募集TGF的“运动受体”？沿微管（MTS）向细胞核的货物运输（即，T？rs，Smads）的IC转运。此外，我们已经确定了KM23/LC7/ROBL/DYNLRB家族的另一个相关成员，称为KM23-2。尽管这种同工型似乎具有类似于KM23-1的特征，但KM23-2似乎更具体地调节SMAD3，而KM23-1的SMAD2调节相反。在新应用程序中将探索与KM23-1/2电动机LCS相关的SMADS2/3差异调节和运输的机制，包括KM23如何影响下游TGF？回答。在最后一个项目期间，我们还显示蛋白激酶A（PKA）直接磷酸化KM23，并且KM23与Dynein中间链（DIC）结合所必需。 TGF中KM23的PKA磷酸化的基础机制？还将检查信号传导和SMAD2/3贩运。其他研究涉及有关这些贩运事件如何改变的人类卵巢癌细胞（HOCC）的研究涉及的，已知以表达改变KM23-1的形式的变化。作为最后一个项目期间的一部分，我们确定了42％的卵巢癌患者组织中的KM23-1改变，在正常组织中未检测到KM23的改变。卵巢癌的如此高的变化率表明，KM23可能在TGF2耐药性或该疾病的肿瘤进展中起重要作用。在新应用中，我们将检查击倒KM23或在关键磷酸化位点突变的影响，对SMAD运输，Smad特异性TGF的影响？反应以及人类癌细胞的恶性表型以及体内肿瘤进展。